"Scaling up bacterial minicell production" This project is focused around scaling up bacterial minicell production to meet projected market demands of any of Vaxiion minicell-based products. We propose to construct a panel of minicell producing strains from E. coli, Salmonella spp, Shigella spp, and Campylobacter using a single genetic approach that has been optimized to maximize minicell productions across the panel. The first aim of this proposal is to identify exactly which genes within the highly conserved ftsQAZ need to be over expressed to obtain maximum minicell production. The second aim of this proposal is to introduce, using the same procedure and materials in parallel, the gene combination identified in the previous effort on to the chromosome of each of the organisms listed. These strains will then be tested for production capacity in growth conditions optimized in small scale (<10L) fermentation runs on an experimental basis in conjunction with a CMO that will eventually be handling all future large scale (<100L) production. PUBLIC HEALTH RELEVANCE: "Scaling up bacterial minicell production" Vaxiion Therapeutics, Inc. Is focused on brining several proprietary bacterial minicell-based products to market. Without going into much detail here, Vaxiion has positioned the bacterial minicell platform technology to be used in vaccine development, targeted delivery of cancer therapeutics, and early stage in vivo targeted tumor bio-imaging. While all three programs are making progress in their respective stages of preclinical development, there remains one vital issue that all of these would be products have in common that must be addressed if minicells are ever to be brought to market. The issue is the ability of Vaxiion to scale up both production and purification to meet projected demand for all of these products. The funds, if awarded for this Phase I SBIR grant proposal will be used to address production, while Phase II would focus primarily on large scale production and purification. [unreadable] [unreadable] [unreadable]